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High Quality Liquid Culture: Step by Step

Good LC, Good Instructions



High Quality Liquid Culture: Step by Step
  • Clean jars with soap and water
  • Soak jars in bleach water for 25 minutes
  • Rinse out well
  • Bring distilled water to boil. Turn off heat
  • Add: 1.5tea/1.5tea malt/dextrose per quart of water (tried higher and lower ratios, this seems to work well)
  • Stir until dissolved. It can take a while for the malt to dissolve, be patient
  • Filter through double coffee filters 3 times. (you can put filters in a funnel or strainer and pour solution through)
  • Fill each quart jar to 600ml (my general rule is 2/3 full for any LC container), make sure liquid is still hot (prevents jar breakage in pc)
  • Put stir bars in
  • Put lids on loose, so nutritious steam doesn't permeate the tyvek as much
  • Put on lid, then tyvek, then ring, then aluminum foil and crimp it around the edges.
  • Put jars in and put the lid on the pc.
  • Put stove temp on med for about 15 mins so the jars can heat without breaking and then turn it up to hot to get the pressure to rise to where you want it.
  • Pressure cook these jars at 14-15psi for 15 minutes (start timer once 14-15psi is reached). Going longer than 15 minutes will ensure better sterilization, but will give you more carmelization. I don't like carmelization, some people don't care. It's up to you. I haven't had a contam yet at this time so....
  • Remove from heat and allow pressure cooker to de-pressurize naturally.
  • Open the pressure cooker and tighten lids as soon as pressure is back to normal. Remove the jars and immediately remove the aluminum foil and wipe off the water on top with a paper towel.
  • Cover lids with a coffee filter dust cover. Held in place with a rubber band.
  • Don't mind the small particles that have formed during the PC. These are good to give the myc something to get growing on.
  • Allow jars to cool for 12-16 hours.
  • Inoculate with 1cc or more of colonized liquid culture or spore solution. Can also inoculate with tissue culture, agar piece, or colonized grain.
  • When you inoculate with spores, it can take up to a week for the spores to germinate and begin to grow.
  • Incubate at 80-85 degrees.
  • After 24 hours, Stir on magnetic stirrer very slow (zero vortex)
  • Stir 24/7, only creating a vortex once or twice daily (for about 3 mins).Having a powerful magnetic stirrer is key to really breaking up the myc well. Some strains might need more recovery time after being vortexed. You might wanna let them sit unstirred for a few hours after vortexing.
  • Colonize jar so mycelium cloud (when allowed to settle without spinning) is about 40-50 percent of liquid.
  • Ready to inoculate spawn/substrate or store in fridge.  I am unsure how long this will stay viable stored in the fridge. You need to keep the fridge in about the 36-40 F range. If your fridge swings to freezing temps easily this can harm or kill the LC. Warm swings will let it grow and get too thick. If you want to play it safe you could make a new LC using this one as inoculant if you've been storing your LC for a while and don't think it will be good for much longer.

Tips:

  • It is advisable to test your LC on a small batch of grain or BRF before committing it to larger endeavors. This way you can visually inspect for contams. Can't see contams easily in LC.
  • Use Light Dry Malt (extra light even better)and Dextrose for this. Both can be found at any beer brewing shop or ordered online from a brewing store. They are cheap and using this is superior to honey or Karo.
  • Use a stir bar and stirring plate. The mycelium gets thick with malt/dex LCs. Without a stir plate it might get too thick to suck into a syringe. Also, you really can't risk shaking the LC and getting the tyvek wet, as you would have to do without a stir plate. This can wick in contaminates.
  • I have found the X shaped stir bars to be easy to work with. They are easier to get off of the side of the jar and spinning in the middle.
  • Have 16 or 14 gauge needles. Any smaller and you will have a very hard time sucking up the mycelium.
  • Use a pressure cooker with a gauge. The Presto 23 qt is a good one for the money. Keep it just under 15 psi (if it has a pressure regulating weight that allows steam to escape) and you won't lose much water inside. This is more important when doing long PC runs with spawnbags or grain jars.
  • I like lids with four 1/4" holes for gas exchange on one side and another hole covered with high temp RTV port sealant on the other side. Holes can be made with a nail and then a phillips head screwdriver. It goes lid->tyvek->ring->foil. The port is a self healing injection site. Also,when you shoot spores or LC into your sugar water you can find your hole easy by feeling for the bump on top of the lid caused by the sealant. You then cover the hole made in the tyvek with micropore tape (or any tape that sticks well, micropore does great for me). When your LC is done you can take off the ring and remove the tyvek and then put the ring back on. Do this in a glove box. Then, while holding glove covered, alcohol swabbed fingers over your air ports, you shake up the mycelium to distribute it throughout the liquid and then insert your needle into the self healing port to suck up your LC. No LC will be leaking out as you tilt the jar towards the needle because of the self healing port. Throw the tyvek back on when you're done. Also, using a double layer of micropore tape over the holes will work as well as tyvek. 
  • After storing LC in the fridge, I would take it out and leave it at room temp for 1-2 days before using it. This will get it growing again and make it less likely to become too shocked to grow after sucking it and dispensing it from syringes.
  • Along with the 16 or 14g needles, get yourself some 60-140cc syringes that you can use to inoculate spawn bags or multiple jars. Look online for farm equipment suppliers. They'll have em. Use 30-300cc's per spawn bag. Try the Poo with Extras Tek .
  • If you can find a good 1/2 gallon or 1 gallon glass jug, use it! Nothing like plentiful LC. You will absolutely need a powerful Magnetic stirrer if using a batch this size though. You might have to compromise on the lid design on one of these. I've used a 1/2 gal jug with a small mouth and just put tyvek over the mouth and kept it in place with zip ties and rubber bands (no lid). Then I would pour the solution into one of my quart jars with an injection port when it was time to suck into syringes.
  • Let me know if anything I've said is unclear or anything is missing or needs further elaboration. I want this to be easy to understand.

-MasFina

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